RESUMEN
Few techniques can assess phenotype and fate for the same cell simultaneously. Most of the current protocols used to characterize phenotype, although able to generate large datasets, necessitate the destruction of the cell of interest, making it impossible to assess its functional fate. Heterogeneous biological differentiating systems like hematopoiesis are therefore difficult to describe. Building on cell division tracking dyes, we further developed a protocol to simultaneously determine kinship, division number, and differentiation status for many single hematopoietic progenitors. This protocol allows the assessment of the ex vivo differentiation potential of murine and human hematopoietic progenitors, isolated from various biological sources. Moreover, as it is based on flow cytometry and a limited number of reagents, it can quickly generate a large amount of data, at the single-cell level, in a relatively inexpensive manner. We also provide the analytical pipeline for single-cell analysis, combined with a robust statistical framework. As this protocol allows the linking of cell division and differentiation at the single-cell level, it can be used to quantitatively assess symmetric and asymmetric fate commitment, the balance between self-renewal and differentiation, and the number of divisions for a given commitment fate. Altogether, this protocol can be used in experimental designs aiming to unravel the biological differences between hematopoietic progenitors, from a single-cell perspective.
Asunto(s)
Hematopoyesis , Células Madre Hematopoyéticas , Ratones , Humanos , Animales , Citometría de Flujo/métodos , Diferenciación Celular , División Celular , FenotipoRESUMEN
Misinformation threatens our societies, but little is known about how the production of news by unreliable sources relates to supply and demand dynamics. We exploit the burst of news production triggered by the COVID-19 outbreak through an Italian database partially annotated for questionable sources. We compare news supply with news demand, as captured by Google Trends data. We identify the Granger causal relationships between supply and demand for the most searched keywords, quantifying the inertial behaviour of the news supply. Focusing on COVID-19 news, we find that questionable sources are more sensitive than general news production to people's interests, especially when news supply and demand mismatched. We introduce an index assessing the level of questionable news production solely based on the available volumes of news and searches. We contend that these results can be a powerful asset in informing campaigns against disinformation and providing news outlets and institutions with potentially relevant strategies.
Asunto(s)
COVID-19 , COVID-19/epidemiología , Comunicación , Brotes de Enfermedades , Humanos , Italia/epidemiología , Medios de Comunicación de MasasRESUMEN
High-throughput single-cell methods have uncovered substantial heterogeneity in the pool of hematopoietic stem and progenitor cells (HSPCs), but how much instruction is inherited by offspring from their heterogeneous ancestors remains unanswered. Using a method that enables simultaneous determination of common ancestor, division number, and differentiation status of a large collection of single cells, our data revealed that murine cells that derived from a common ancestor had significant similarities in their division progression and differentiation outcomes. Although each family diversifies, the overall collection of cell types observed is composed of homogeneous families. Heterogeneity between families could be explained, in part, by differences in ancestral expression of cell surface markers. Our analyses demonstrate that fate decisions of cells are largely inherited from ancestor cells, indicating the importance of common ancestor effects. These results may have ramifications for bone marrow transplantation and leukemia, where substantial heterogeneity in HSPC behavior is observed.
Asunto(s)
Diferenciación Celular , Proliferación Celular , Células Madre Hematopoyéticas/fisiología , Animales , Médula Ósea , Células de la Médula Ósea , Células Cultivadas , Células Madre Hematopoyéticas/clasificación , Ratones , Ratones Endogámicos C57BLRESUMEN
Lymphocytes are the central actors in adaptive immune responses. When challenged with antigen, a small number of B and T cells have a cognate receptor capable of recognising and responding to the insult. These cells proliferate, building an exponentially growing, differentiating clone army to fight off the threat, before ceasing to divide and dying over a period of weeks, leaving in their wake memory cells that are primed to rapidly respond to any repeated infection. Due to the non-linearity of lymphocyte population dynamics, mathematical models are needed to interrogate data from experimental studies. Due to lack of evidence to the contrary and appealing to arguments based on Occam's Razor, in these models newly born progeny are typically assumed to behave independently of their predecessors. Recent experimental studies, however, challenge that assumption, making clear that there is substantial inheritance of timed fate changes from each cell by its offspring, calling for a revision to the existing mathematical modelling paradigms used for information extraction. By assessing long-term live-cell imaging of stimulated murine B and T cells in vitro, we distilled the key phenomena of these within-family inheritances and used them to develop a new mathematical model, Cyton2, that encapsulates them. We establish the model's consistency with these newly observed fine-grained features. Two natural concerns for any model that includes familial correlations would be that it is overparameterised or computationally inefficient in data fitting, but neither is the case for Cyton2. We demonstrate Cyton2's utility by challenging it with high-throughput flow cytometry data, which confirms the robustness of its parameter estimation as well as its ability to extract biological meaning from complex mixed stimulation experiments. Cyton2, therefore, offers an alternate mathematical model, one that is, more aligned to experimental observation, for drawing inferences on lymphocyte population dynamics.
RESUMEN
When COVID-19 pandemic spread in Europe, governments imposed unprecedented confinement measures with mostly unknown repercussions on contemporary societies. In some cases, a considerable drop in energy consumption was observed, anticipating a scenario of sizable low-cost energy generation, from renewable sources, expected only for years later. In this paper, the impact of governmental restrictions on electrical load, generation and transmission was investigated in 16 European countries. Using the indices provided by the Oxford COVID-19 Government Response Tracker, precise restriction types were found to correlate with the load drop. Then the European grid was analysed to assess how the load drop was balanced by the change in generation and transmission patterns. The same restriction period from 2020 was compared to previous years, accounting for yearly variability with ad hoc statistical technique. As a result, generation was found to be heavily impacted in most countries with significant load drop. Overall, generation from nuclear, and fossil coal and gas sources was reduced, in favour of renewables and, in some countries, fossil gas. Moreover, intermittent renewables generation increased in most countries without indicating an exceptional amount of curtailments. Finally, the European grid helped balance those changes with an increase in both energy exports and imports, with some net exporting countries becoming net importers, notably Germany, and vice versa. Together, these findings show the far reaching implications of the COVID-19 crisis, and contribute to the understanding and planning of higher renewables share scenarios, which will become more prevalent in the battle against climate change.
RESUMEN
The generation of cellular heterogeneity is an essential feature of immune responses. Understanding the heritability and asymmetry of phenotypic changes throughout this process requires determination of clonal-level contributions to fate selection. Evaluating intraclonal and interclonal heterogeneity and the influence of distinct fate determinants in large numbers of cell lineages, however, is usually laborious, requiring familial tracing and fate mapping. In this study, we introduce a novel, accessible, high-throughput method for measuring familial fate changes with accompanying statistical tools for testing hypotheses. The method combines multiplexing of division tracking dyes with detection of phenotypic markers to reveal clonal lineage properties. We illustrate the method by studying in vitro-activated mouse CD8+ T cell cultures, reporting division and phenotypic changes at the level of families. This approach has broad utility as it is flexible and adaptable to many cell types and to modifications of in vitro, and potentially in vivo, fate monitoring systems.